LOCUS       sequence-402996-        5180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARL5B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5180)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5180)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pET3-ARL5B-His6
FEATURES             Location/Qualifiers
     source          1..5180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             708..725
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(779..797)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      793..840
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1206..1211,1212..1228,1229..1234))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1261..1279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1347..1451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1452..1520,1521..2312)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1670..1689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2483..3071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2972..2991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3225..3242
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3257..3399
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3325..3344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3485..3507
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3501..3692)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5150..5169
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
       61 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      121 attttgttta actttaagaa ggagatatac atatggggct gatcttcgcc aaactgtgga
      181 gcctcttctg taaccaagaa cacaaagtaa ttatagtggg actggataat gcagggaaaa
      241 ccaccattct ttaccaattc ttaatgaatg aagtggttca tacttctcca accataggaa
      301 gcaatgttga agaaatagtt gtgaagaaca ctcattttct tatgtgggat attggtggtc
      361 aggagtctct gcgatcatcc tggaacacat attactcaaa tacagagttc atcattcttg
      421 ttgttgatag cattgacagg gaacgactag ctattacaaa agaagaatta tacagaatgt
      481 tggctcatga ggatttacgg aaggctgcag tccttatctt tgcaaataaa caggatatga
      541 aagggtgtat gacagcagct gaaatctcga aatacctcac ccttagttca attaaggatc
      601 atccatggca cattcaatcc tgctgtgctc tcacaggaga agggttatgc caaggtctag
      661 agtggatgac ctcccggatt ggtgtgagag gcagcggcag cggcagccat caccatcacc
      721 atcactaagg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
      781 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      841 ctgaaaggag gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta
      901 gtcgatagtg gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga
      961 cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac
     1021 gccatagtga ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac
     1081 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
     1141 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
     1201 accgcattaa agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg
     1261 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1321 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1381 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1441 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1501 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1561 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1621 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1681 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1741 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1801 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1861 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1921 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1981 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2041 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2101 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2161 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2221 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2281 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3121 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3181 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3241 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3301 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3361 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3421 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3481 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3541 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3601 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3661 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3721 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3781 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3841 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3901 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3961 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4021 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4081 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4141 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     4201 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     4261 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4321 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4381 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4441 cggcataaat cgccgtgacg atcagcggtc cagtgatcga agttaggctg gtaagagccg
     4501 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4561 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4621 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4681 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4741 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4801 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4861 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4921 tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca
     4981 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     5041 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5101 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5161 cgatataggc gccagcaacc
//
